title: Cloning to crystallization of dihydrodipicolinate synthase from the intracellular pathogen Legionella pneumophila.
abstract: Dihydrodipicolinate synthase (DHDPS) catalyses the rate-limiting step in the biosynthesis of meso-diaminopimelate and lysine. Here, the cloning, expression, purification and crystallization of DHDPS from the intracellular pathogen Legionella pneumophila are described. Crystals grown in the presence of high-molecular-weight PEG precipitant and magnesium chloride were found to diffract beyond 1.65 Å resolution. The crystal lattice belonged to the hexagonal space group P6₁22, with unit-cell parameters a=b=89.31, c=290.18 Å, and contained two molecules in the asymmetric unit. The crystal structure was determined by molecular replacement using a single chain of Pseudomonas aeruginosa DHDPS as the search model.
authors.affiliation: Department of Biochemistry, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, VIC 3086, Australia.
authors.forename: Tanzeela
authors.initials: T
authors.lastname: Siddiqui
authors.forename: Jason J
authors.initials: JJ
authors.lastname: Paxman
authors.forename: Con
authors.initials: C
authors.lastname: Dogovski
authors.forename: Santosh
authors.initials: S
authors.lastname: Panjikar
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Perugini
mesh_terms: D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D006836:Hydro-Lyases / Q000737:chemistry* / Q000302:isolation & purification; D042541:Intracellular Space / Q000469:parasitology*; D016952:Legionella pneumophila / Q000201:enzymology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D011994:Recombinant Proteins; D006836:Hydro-Lyases; C020040:4-hydroxy-tetrahydrodipicolinate synthase
keywords: Legionella pneumophila; dihydrodipicolinate synthase